Int. J. Pharm. Sci. Rev. Res., 25(2), Mar – Apr 2014; Article No. 07, Pages: 33-36
ISSN 0976 – 044X
Research Article
Phytochemical Analysis of Methanolic Extract of Cuscuta reflexa Grown on
Cassia fistula and Ficus benghalensis by GC-MS
Neetu Bais*, Arun Kakkar
Govt. Model Science College, Jabalpur (M.P), India.
*Corresponding author’s E-mail: baisneetu@yahoo.co.in
Accepted on: 15-01-2014; Finalized on: 31-03-2014.
ABSTRACT
The objective of present study was to analyze the phytochemicals present in the Cuscuta reflexa parasite grown on two different
plant species. A comparative GC-MS analysis was done on methanol extract of Cuscuta reflexa grown on Cassia fistula and Ficus
benghalensis. This analyses revealed that benzofuran 2, 3, dihydro-, 2-methoxy-4-vinylphenol and 2-propenoic acid, 3-(4hydroxyphenyl)-methyl ester were present in both Cuscuta reflexa grown on Cassia fistula and Cuscuta reflexa grown on Ficus
benghalensis. It was found that urea N, N’-bis(1-methylethyl)-, 6-Chloro-4-phenyl-2-propylquinoline and benzo[h]quinoline 2, 4dimethyl- were present only in Cuscuta reflexa grown on Cassia fistula while propanedioic acid, propyl, 2-[2-thienyl]-4-acetyl
quinoline and L-alanine-4-nitroanilide were found to be present only in Cuscuta reflexa grown on Ficus benghalensis. This variation
in phytochemicals present in Cuscuta reflexa parasite grown on Cassia fistula and Ficus benghalensis confirms that chemical
constituents of parasite depends on nature of host.
Keywords: Cuscuta reflexa, Cassia fistula, Ficus benghalensis, GC-MS analysis.
INTRODUCTION
M
edicinal plants have been used since ages in
traditional medicines due to their therapeutic
potential and the search in medicinal plants
have led to the discovery of novel drug candidates used
against various diseases.1 The treatment of various
diseases with indigenous medicinal plants generates
considerable health and economic benefits. Traditional
knowledge in this regard has been conserved for
generations in different tribal communities in several
parts of India.2
Knowledge of potential chemical constituents is
important for the discovery of therapeutic agents, which
may be of great value as the source of economical
phytocompounds for the synthesis of complex chemical
substances.
Cuscuta reflexa is rootless, leafless perennial parasitic
twining herb of convolulaceae family, commonly known
as amarbel or dodder. It has no chlorophyll and cannot
make its own food by photosynthesis. It spread from one
host plant to another, and on each victim, they twin and
cling tightly with special branching organs called
haustorium. Haustorium penetrate the host and connect
to the host xylem as well as to the host phloem and
absorb from it both water and elaborated food stuff. 3 It is
believed that the parasitic herbs extract healthy and
potential sap from host plant and if their host plants are
medicinal plants then these parasitic herbs show many
similar properties to host plants. Cuscuta species feeding
on commonly used medicinal herbs are given special
attention by traditional healers.
Cuscuta reflexa is the valuable medicinal herb. Stem of
this plant is antibacterial and used externally to treat itch
and internally in fever. 4 It is useful in treatment of
androgen induced alopecia. 5
It also gives anti
inflammatory and anti cancer activity.6 The aqueous and
alcoholic extract of C. reflexa has diuretic activity.7 The
crude water extract of the C. reflexa also shows the anti
HIV activity. 8
MATERIALS AND METHODS
Collection of plant material
Cuscuta stem were collected from the tree of Cassia
fistula and Ficus benghalensis tree near village areas of
Gokulpur in Jabalpur Madhya Pradesh, India in the month
of September and November 2010 respectively. Collected
plant material was authenticated by Dr. Indu Gupta
Botany Dept. Govt. Model Science College Jabalpur (M.P),
India. Stems were washed thoroughly with water.
Immense care was taken to avoid the mixing of host plant
with that of targeted Cuscuta stem. Stems of Cuscuta
were cleaned and completely separated from the stems
of host plant.
Solvent extraction
Thoroughly washed stems of Cuscuta reflexa from both
the host trees were shade dried for 15 days and the
powdered in the grinder. The shade dried powder was
extracted with petroleum ether, ethyl acetate, methanol
and water in increasing polarity. The extracts were filter
with Whatman’s filter paper. Filtrates were concentrated
under reduced pressure and preserved at 5oC in dark air
tight bottles
Sample preparation for GC-MS analysis
50 µl of sample was dissolve in 2ml of methanol and kept
in ultrasonic bath for 15 min and centrifuged for 10 min at
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Int. J. Pharm. Sci. Rev. Res., 25(2), Mar – Apr 2014; Article No. 07, Pages: 33-36
6000 rpm and supernatant was injected in GC-MS for
analysis.
GC-MS analysis
GC-MS of methanol extract was performed using Agilent
7890A. Compounds were separated on Agilent 1909S433: 2065.49541 HP -5MS. 5% phenyl methyl silox column
(30m x 250µm x 0.25µm). Oven temperature was
programmed as follows: isothermal temperature of 50o C
for 2 min then increased to 150oC at the rate of 5oC /min
o
and held for 1.75 min then increased to 280 C at the rate
of 8oC /min and kept constant for 5min. The run time was
45 min. ionization of sample components were
performed on EI mode (70 eV). The carrier gas was helium
at 1.0ml/min flow rate. 0.5 ml of sample was injected in
split mode of 20:1.The mass spectrum scan range was set
at 29.0 to 500(m/z).
Identification of compounds
Interpretation of mass spectrum of GC-MS was done
using the database of National Institute Standard and
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Technology (NIST). The mass spectrum of phytochemicals
was compared with the spectrum of known compounds
stored in the NIST library.
RESULTS
Phytochemicals present in methanol extract in Cuscuta
reflexa grown on C. fistula and F. benghalesis are
summarized in table 1 and table 2.
Figure 1 represents the GC chromatogram of methanol
extract of C. reflexa grown on C. fistula. It shows many
peaks out of which six major peaks were characterized
and identified on comparison of mass spectra with NIST
library. Fig 3 represents the mass spectrum of
components present in C. reflexa grown on C. fistula.
Fig 2 shows the GC chromatogram of methanol extract of
C. reflexa grown on F. benghalensis. Out of many peaks
six important peaks were characterized. Fig 4 shows the
mass spectrum of phytochemicals identified in C. reflexa
grown on F. benghalensis.
Table 1: Phytochemicals identified in methanol extract of C. reflexa grown on C. fistula
S.No
RT
Name of Compound
MF
MW
Total peak %
1.
15.354
Benzofuran,2,3-dihydro-
C8H8O
120.14
38.024
2.
18.045
2-Methoxy-4-vinylphenol
C9H10O
150.17
3.473
3.
19.682
Urea, N, N’- bis(1-methylethyl)-
C7H16N2O
144.21
0.866
4.
28.583
2-Propenoic acid, 3-(4-hydroxyphenyl)-, methyl ester
C10H10O3
178.18
7.585
5.
29.267
6-Chloro-4-phenyl-2-propylquinoline
C18H16ClN
281.77
7.445
6.
31.547
Benzo[h]quinoline,2, 4-dimethyl-
C15H13N
207.27
1.234
Table 2: Phytochemicals identified in methanol extract of C. reflexa grown on F. benghalensis
S.No
RT
Name of Compound
MF
MW
Total peak %
1.
15.354
Benzofuran,2,3-dihydro-
C8H8O
120.14
24.971
2.
18.045
2-Methoxy-4-vinylphenol
C9H10O
150.17
2.400
3.
19.682
Propanedioic acid, propyl
C6H10O4
146.14
8.021
4.
28.583
2-Propenoic acid, 3-(4-hydroxyphenyl)-, methyl ester
C10H10O3
178.18
3.809
5.
29.267
2-[2-Thienyl]-4-acetyl quinoline
C15H11NOS
253.31
15.241
6.
31.547
L-Alanine-4-nitroanilide
C9H11N3O3
209.2
1.733
Figure 1: GC-MS Chromatogram of methanol extract of C. reflexa grown on C. fistula
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Int. J. Pharm. Sci. Rev. Res., 25(2), Mar – Apr 2014; Article No. 07, Pages: 33-36
ISSN 0976 – 044X
Figure 2: GC-MS Chromatogram of methanol of C. reflexa grown on F. benghalensis
Figure 3: Mass spectrum of phytochemicals identified in C. reflexa grown on C.fistula
Figure 4: Mass spectrum of phytochemicals identified in C. reflexa grown on F. benghalensis
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Int. J. Pharm. Sci. Rev. Res., 25(2), Mar – Apr 2014; Article No. 07, Pages: 33-36
Retention times of phytochemicals identified in methanol
extract of C. reflexa grown on C. fistula were found to be
15.354, 18.045, 19.682, 28.583, 29.267 and 31.547.
Phytochemicals identified in C. reflexa grown on C. fistula
were benzofuran 2, 3-dihydro- (38.024%), 2-methoxy-4vinylphenol (3.473%), Urea, N, N’- bis (1-methylethyl)(0.866%), 2-propenoic acid, 3-(4-hydroxyphenyl)-,methyl
ester (7.585%), 6-Chloro-4-phenyl-2-propylquinoline
(7.445%), benzo[h]quinoline,2, 4-dimethyl- (1.234%).
From GC-MS analysis of methanol extract of C. reflexa
grown on F. benghalensis six compounds were identified.
Retention times of these compounds were found to be
15.354, 18.046, 25.479, 28.582, 29.240 and 32.998. The
phytochemicals identified were benzofuran 2 ,3-dihydro(24.971%),
2-methoxy-4-vinylphenol
(2.400%),
propanedioic acid, propyl (8.021%), 2-propenoic acid,3-(4hydroxyphenyl)-methyl ester (3.809%), 2-[2-Thienyl]-4acetyl quinoline (15.241%), L-alanine-4-nitroanilide
(1.733%).
concluded that variation in phytochemicals in C. reflexa is
host dependent. Further study need to be undertaken to
investigate the biological activity and other
phytochemicals present in C. reflexa grown on C .fistula
and F. benghalensis.
REFERENCES
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Sermakkani M, Thangapandian V. GC-MS analysis of Cassia
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Plant used by the Thakar Tribes of Raigad district,
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Kaur A. Cuscuta reflexa Roxb. A parasitic plant in Ayurved.
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Pal DK, Mandal M, Senthil Kumar GP, Padhiari A.
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Pandit S, Chauhan NS, Dixit VK. Effect of Cuscuta reflexa
ROXB on androgen induces alopecia. J. Cosmet. Dermatol. ,
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Suresh V, Sruthi V, Padmaja B, Asha VV. In vitro anti
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Sharma S, Hullati KK, Prasanna SM, Kuppast IJ, Sharma P.
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DISCUSSION
From table 1 and Table 2 it was revealed that benzofuran
2, 3, dihydro- , 2-methoxy-4-vinylphenol and 2-propenoic
acid, 3-(4-hydroxyphenyl)-methyl ester was common to
both C. reflexa grown on C. fistula and C. reflexa grown on
F. benghalensis. It was found that urea N, N’-bis(1methylethyl)-, 6-Chloro-4-phenyl-2-propylquinoline and
benzo[h]quinoline,2, 4-dimethyl- were present only in C.
reflexa grown on C. fistula while Propanedioic acid,
propyl, 2-[2-Thienyl]-4-acetyl quinoline and L-alanine-4nitroanilide were found to be present only in C. reflexa
grown on F. benghalensis.
Benzo[h]quinolines are found to have wound healing,
antibacterial, DNA binding and antioxidant property.9
Quinolines and its derivatives possess anticancer,
antimicrobial, anticonvulsant, anti-inflammatory and
cardiovascular activity.10 2-Propenoic acid, 3-(4hydroxyphenyl)-, methyl ester is the derivative of pcoumaric acid which is worked out for its anti oxidant and
antimicrobial activity.11
CONCLUSION
It is revealed from this study that C. reflexa from both the
host trees is rich in secondary metabolites which possess
wide range of biological activities. Different compounds
are present in C. reflexa on two different host thus it is
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10. Kumar S, Bawa S, Gupta H. Biological activities of quinoline
derivatives. Mini Rev Med Chem., 9(14),2009, 1648-54.
11. Stojkovic D, Petrovic J, Sokovic M, Glamoclija J, KukicMarkovic J, Petrovic S. In situ antioxidant and antimicrobial
activities of naturally occurring Caffeic acid, P-coumaric
acid and rutin using food systems. J Sci Agric, 93(13), 2013,
3205-3208.
Source of Support: Nil, Conflict of Interest: None.
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